Reid Ruth, Samaras Shivanthi, The Development and Evaluation of A Multiplex Real-Time PCR Assay for the Detection of ESBL Genes in Urinary Tract Infections, International Journal of Clinical Microbiology, Volume 1, Issue 1, 2018, Pages 16-24, ISSN 2690-4721, https://doi.org/10.14302/issn.2690-4721.ijcm-18-2217.
(https://oap-researcharticles.org/ijcm/article/808)
Abstract: Background Overuse of beta-lactam antibiotics has lead to selection for extended-spectrum β-lactamase (ESBL) producing Enterobacteriaceae, a major cause of antibiotic resistant urinary tract infections (UTIs). Standard detection methods are time-consuming, with disputed accuracy. This study describes a novel real-time PCR method to detect CTX-M, SHV, OXA and TEM. Methods 179 Enterobacteriaceae isolates from UTIs were collected from the Leicester Royal Infirmary, UK. A multiplex Plexor®-based real-time PCR assay detected ESBLs using their specific amplicon melting temperature, during each cycle, removing the need for a melt-curve analysis. Validation was achieved by end-point PCR and disk diffusion. Results The method was able to produce rapid and accurate results, achieving a sensitivity and specificity of 94.9% and 72% respectively, and the assay can differentiate between the different ESBL genes, with ease. Conclusions With further investigation, a Plexor®-based assay could form the basis of a high-throughput kit that health services could use to detect ESBLs or other antibiotic resistance genes.
Keywords: Extended-spectrum beta-lactamases; urinary tract infections; Real-Time PCR; rapid diagnostics; antibiotic resistance.